International Journal of Smart Grid and Clean Energy

Effect of Fermentation on Extraction of Essential Oils on

Fennel Plants (Eupatorium capillifolium)
Andi Nursantia, Annisa Permatasarib, FaradisaAninditab, Gustini Syachbirinb*
a
Departement of Chemistry, BogorAgriculture University, Dramaga, Bogor 16680, Indonesia
b
Departement of Chemistry, BogorAgriculture University, Dramaga, Bogor 16680, Indonesia

Abstract
Fennel plants species Eupatorium capillifolium contains essential oils. The research was conducted to determine the
effect of Fermentation on Extraction of Essential Oils on Fennel (E. Capillifolium) and chemical compound group
contained in essential oil. Methods used dried, fermentation using water, Efective Microorgaisms 4 (EM4) and
Aspergilus, sp as well as extraction of essential oils using steam distillation. The result of fermentation method has
influence yield of essential oil of E. capillifolium that is produced that increase two times compared with dry method.
The best fermentation method is a mild fermentation method by adding water to the sample yielding the highest yield.
The active compounds contained in the essential oil of E. capillifolium are α-phellandrene (22.57%), trans-
caryophyliene (10.70%), 2-butanone (10.04%), thymyl methyl ether (6.56%), germacrene (5.26%) , and cyclohexane
(4.77%).
Keywords: essential oils, Eupatorium capillifolium, fermentation

1. Introduction

The fennel plants (Eupatorium capillifolium) are native to North America and also grow in the
southeastern United States [1]. This type then goes into different countries, so it also exists in Burma,
India, Nepal, and Malaysia. In Indonesia, E. capillifolium is known as bungo china in West Sumatra and
is used only as an ornamental plant and a hedge plant [2]. E. capillifolium species contain several lactate
compounds, flavonoids, phenolic compounds and acetylenic acids, triterpenes and alkaloids. This
compound has cytotoxic, antitumoral, antimicrobial, antioxidant and antiinflammatory activity [3]. Group
terpenoid compounds is the main fraction of essential oil, this substance is the cause of fragrant, fragrant
or distinctive smells in many plants. Economically these compounds are important as the basis of natural
fragrance and also for spices as well as flavor compounds in the food industry [4].
The study of essential oil E. capillifolium from Indonesia has never been done. [5] So far the report of
essential oil of E. capillifolium was reported using E. capillifolium plants from Missisipi, USA. The
essential oil obtained is yellow with a strong aromatic odor with 0.4% reduction. The main components of
oil were found as methyl etherol (methyl timolol) (36.3%), 2,5-dimethoxy-p-cymene (20.8%) and
myrcene (15.7%). [6] Essential oil on patchouli produced with several techniques such as distillation,
extraction and fermentation techniques. The yield of patchouli oil from dried leaves obtained by
distillation technique is 0.73%, the extraction technique is 3.56% while the fermentation technique is
6.22%. [7] The content of fennel oil on fennel seeds is 0.607% and on fennel leaves is 0.27% with
refining time for 4 hours.
Based on the above, then the research was conducted to determine the effect of fermentation on
extraction of essential oils on fennel leaf plants (E. capillifolium) and chemical compounds contained in
essential oil.

1. Experimental Procedure
1.1 Materials
2 International Journal of Smart Grid and Clean Energy, vol. 2, no. X, Month 2013

E. capillifolium were collected from Bogor, Indonesia, distilled water , anhydrous

Na2SO4, Efective Microorgaisms (EM4) and Aspergilus niger was obtained from BBP
laboratory microbiology, Bogor Indonesia.

1.2 Preparation of E. capillifolium leafs fermentation and non fermentation

Fennel plants are separated by leaves and stems, partially dried and others are
chopped to be fermented. Fermentation is done in 3 types, the first is done by spinning
distilled water until wet evenly on the sample, the second is done by spinning distilled
water + Efective Microorgaisms (EM4), and third by sprinkling distilled water +
Aspergilus niger. Then covered in a black plastic container until no air is allowed to
stand for 10 hours. The dried and fermented sample is ready to distilled its oil.

1.3 Preparation of E. capillifolium to steam distilled

Three kilograms of sample were put into pot , then added 5 Liter of distilled water.
distillation process carried out for 3 hours. After the distillation process is complete the
oil is collected and anhydrous Na2SO4 is added. The extract was stored in refrigerator
until required for use. The same procedure was followed for all varieties fermented and
dry of E. capillifolium.

1.4 Preparation of E. capillifolium to identification of component components with GC-

MS
The essential oil samples from fennel leaf were dissolved in 5 mL of isooktana
solvent: toluene (9: 1) then analyzed by Agilent 7890B / 7000C GC-MS Triple
Quadrupole system. The stationary phase of the HP-5MS column (20 m x 0.25 mm x
0.25 μm). Gas carrier helium with a flow rate of 2.25 mL/minute. 2 μL injection volume
with 200:1 split ratio. The column temperature is set to a programmable temperature at
50°C for 1 minute and then raised 20°C to 210° C and held for 5 minute. The compound
essential oil component of fennel leaf is identified by comparing the mass spectrum
with the NIS library.

2. Results and Discussion

3.1 Effect of Fermentation on Extraction of Essential Oils fermentation and non
fermentation
The yield of essential oil is calculated based on the weight of oil obtained from
the distillation process using distilled water. The preliminary treatment of fermentation
has an effect on the yield of essential oil produced. The experimental results showed a
doubling of yield by using distilled water and fermentation method when compared
with dry method (Table 1). According to Guenther [8] essential oils in aromatic plants
are surrounded by oil glands, vessels, oil sacs or granular hair. If the material is left
intact, the essential oil can only be extracted when water vapor succeeds through the
plant tissue and pushes it to the surface. The process of fermentation prior to oil
extraction is well done with the aim of breaking the oil cells [9].
Table 1 Effect of preliminary treatment on essential oil yield
 Authoret al.: Click 3

Volume of Essential The yield of

Preliminary Treatment
Oils Essential Oils
Dried 10.05 ml 0.30%

Fermented distiled water 20 ml 0.60%

Distilled water + Efective 17.6 ml 0.50%

Microorgaisms

Distilled water + Aspergilus niger 18.5 ml 0.50%

3.2 Effect essential oil yield E. capillifolium fermentation and non fermentation
The principle of fermentation in fennel oil isolation is by breaking the cell wall of
the plant by using enzymes contained in microorganisms. The destruction of the cell
wall results in essential oil separate from the leaves and can be extracted more easily
[10]. In the fermentation process occurs a process of biodelignification, so that oil in the
leaf vakuola can be taken.
During the fermentation process, the growing microorganisms will produce some
extracellular enzymes. Cellulase enzyme is an enzyme that has a high enough ability to
hydrolyze cellulose and hemicellulose, so oil molecules bound in leaf vacuoles are
easier to distill. The performance of steam during distillation in order to extract oil
molecules did not experience significant obstacles [11]. The presence of cellulase
enzymes can lead to breakdown of water-insoluble cellulose into water-soluble
components such as selobiose and glucose.
The fermentation process is also carried out using the aid of microorganisms
intentionally added to the sample before being allowed to stand for 10 hours. The
microorganisms used here are Efective Microorganism 4 (EM4) and Aspergilus niger,
the addition of these two microbes aims to increase the yield of essential oils produced.
The experimental results (Table 1) showed no increase or influence of the addition of
microorganisms to the yield of essential oils produced. This is thought to be caused by a
less optimal fermentation time for the growth of microorganisms to produce cellulase
enzymes.

Acknowledgment

This research work is supported by departement of Chemistry, Bogor Agriculture University,

Indonesia. Andi Nursanti would also like to thank the Indonesia Endowment Fund for Education (LPDP)
for their support on his studied.

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There is also the option to include a subheading within the Appendix if you wish.

Authors Information
4 International Journal of Smart Grid and Clean Energy, vol. 2, no. X, Month 2013

Author name Title Membership Contact Author affiliation

Given name Family Prof./A. Prof./ IEEE Member, E-mail/ Affiliation, Address, City and Postcode,
name Dr./S.E.* SM, Fellow** Mobile phone Country

*S.E.—Senior Engineer; ** Member of IEEE—Fellow/Senior member/Member